Probiotics and Antimicrobial Proteins最新文献

In this study, we aimed to develop a protective probiotic coculture to inhibit the growth of Salmonella enterica serovar Typhimurium in the simulated chicken gut environment. Bacterial strains were isolated from the digestive mucosa of broilers and screened in vitro against Salmonella Typhimurium ATCC 14028. A biocompatibility coculture test was performed, which identified two biocompatible strains, Ligilactobacillus salivarius UO.C109 and Ligilactobacillus saerimneri UO.C121 with high inhibitory activity against Salmonella. The cell-free supernatant (CFS) of the selected isolates exhibited dose-dependent effects, and the inhibitory agents were confirmed to be proteinaceous by enzymatic and thermal treatments. Proteome and genome analyses revealed the presence of known bacteriocins in the CFS of L. salivarius UO.C109, but unknown for L. saerimneri UO.C121. The addition of these selected probiotic candidates altered the bacterial community structure, increased the diversity of the chicken gut microbiota challenged with Salmonella, and significantly reduced the abundances of Enterobacteriaceae, Parasutterlla, Phascolarctobacterium, Enterococcus, and Megamonas. It also modulated microbiome production of short-chain fatty acids (SCFAs) with increased levels of acetic and propionic acids after 12 and 24 h of incubation compared to the microbiome challenged with S. Typhimurium. Furthermore, the selected probiotic candidates reduced the adhesion and invasion of Salmonella to Caco-2 cells by 37-39% and 51%, respectively, after 3 h of incubation, compared to the control. These results suggest that the developed coculture probiotic strains has protective activity and could be an effective strategy to control Salmonella infections in poultry.

Probiotics, such as Lacticaseibacillus rhamnosus, are essential to the food industry for their health benefits to the host. The Lcb. rhamnosus strain is susceptible to processing, gastrointestinal, and storage conditions. In this study, Lcb. rhamnosus strains were encapsulated by complex coacervation in a gum arabic/chitosan or gum arabic/trehalose/chitosan and cross-linked with sodium tripolyphosphate. The physicochemical properties (zeta potential, water activity, water content, and hygroscopicity), encapsulation efficiency, and probiotic survival under storage conditions and simulated gastrointestinal fluids were evaluated. The results showed that crosslinking improves the encapsulation efficiency after drying; however, this result was remarkable when trehalose was used as a cryoprotectant. Furthermore, the encapsulation matrix preserved the viability of probiotics during 12 weeks with probiotic counts between 8.7-9.5, 7.5-9.0, and 5.2-7.4 log₁₀ CFU g^-1 at -20, 4, and 20 °C, respectively. After 12 days of digestion in an ex vivo simulator, acetic, butyric, propionic, and lactic acid production changed significantly, compared to free probiotic samples. This work shows that encapsulation by complex coacervation can promote the stability of probiotic bacteria in storage conditions and improve the viability of Lcb. rhamnosus HN001 during consumption so that they can exert their beneficial action in the organism.

Listeria monocytogenes is a significant foodborne pathogen that can form biofilms on various food processing surfaces, thereby enhancing resistance to disinfectants and exacerbating harm to human health. Previous studies have indicated that bacteriocin BM173 exhibits antibacterial and antibiofilm activities. In the current study, our aim was to assess the inhibitory mode of action of sub-inhibitory concentrations (SICs, 1/32 × MIC and 1/16 × MIC) of BM173 on the biofilm formation L. monocytogenes. Crystal violet staining assay revealed that SICs of BM173 significantly inhibit L. monocytogenes biofilm formation. Furthermore, the results of swimming motility assay, plate count, ruthenium red staining, and scanning electron microscopy (SEM) revealed that SICs of BM173 could effectively reduce the movement, cell adhesion, and exopolysaccharide (EPS) production of L. monocytogenes, thereby inhibiting biofilm formation. Real-time quantitative PCR analyses further demonstrated that SICs of BM173 down-regulated the expression of biofilm-associated genes, including those encoding adhesion, virulence factors, and quorum sensing. Additionally, SICs of BM173 effectively reduced the biofilm formation of L. monocytogenes on the surfaces of three food-grade materials (glass, stainless steel, and silicone) at 4 and 25 °C. These outcomes suggest that BM173 holds great potential for development as a promising food preservative for application in the food industry.

Diabetes mellitus type 2 (DM2) is the most common chronic disease worldwide, characterized mainly by increased glucose concentration in the blood and affecting several organs' functionality. The daily consumption of probiotic bacteria can help control diabetes and reduce the damage caused. Cell immobilization techniques are a powerful tool that provides physical cell protection to such probiotic bacteria against gastrointestinal conditions. We suggest that cell immobilization could be a significant vector for delivering a high quantity of viable probiotics to the gut, helping attenuate hyperglycemia in diabetic rats. Seventy male Wistar rats were used in this work. Nicotinamide was administrated via intraperitoneal injection 15 minutes before inducing type 2 diabetes (DM2), followed by a second intraperitoneal injection of streptozotocin to induce DM2. Rats were divided into seven groups. For 45 days, a specific treatment was applied to each group. The group of rats, supplied with immobilized Lactobacillus casei, showed a serum glucose concentration of 137 mg/dL, which was close to the one observed in the groups of healthy rats (117 mg/dL) and rats treated with metformin (155 mg/dL). The diabetic rats without treatment presented a higher serum glucose concentration (461 mg/dL). In the rats treated with immobilized L. casei, there was no biochemical parameter alteration, and the cell morphology of the analyzed tissues was similar to those of the healthy group. The consumption of immobilized L. casei could allow a high quantity of viable probiotics to be delivered to the gut, reducing serum glucose concentration by up to 70% compared to diabetic rats and reducing organ damage caused by diabetes.

This study aimed to investigate the effects of dietary Bacillus subtilis supplementation on gut microbiota diversity, digestive enzyme activity, and Toll-like receptor (TLR) expression in broiler chickens. A total of 240 "817" crossbred broiler chickens were randomly assigned to four groups: control (basal diet, BD), group I (BD + 300 g/d B. subtilis at 1.08 × 107 CFU/kg), group II (BD + 600 g/d B. subtilis at 2.16 × 107 CFU/kg), and group III (BD + 900 g/d B. subtilis at 3.24 × 107 CFU/kg). Gut microbiota analysis revealed significant improvements in the abundance of specific microorganisms in the treatment groups, with distinct variations in the core microorganisms between the groups. Notably, protease activity in the ileum was significantly increased in groups II (22.59%; p < 0.01) and III (14.49%; p < 0.05) compared to that in the control group. Moreover, significant up-regulation of TLR1A and TLR7 expression was observed in jejunum and cecum of the treated groups. Additionally, the TLR1B expression in the ileum was significantly increased. Furthermore, TLR2A and MyD88 transcription levels were significantly elevated in the jejunum, liver, spleen, and kidneys of experimental groups. Modulations in the expression of various TLR's (TLR2B, TLR3, TLR4, TLR15, and TLR21) were also observed in different organs. The spleen and kidney of B. subtilis-supplemented chickens exhibited upregulated expression of the proinflammatory cytokine IL-1β. Dietary supplementation with B. subtilis in broiler chickens improved the gut microbiota diversity and significantly upregulated TLR's expression in various organs. B. subtilis could be a valuable feed additive, contributing to improved disease management and overall health in broiler chickens.

Antimicrobial peptides (AMPs) are the most favorable alternatives in overcoming multidrug resistance, alone or synergistically with conventional antibiotics. Plant-derived AMPs, as cysteine-rich peptides, widely compensate the pharmacokinetic drawbacks of peptide therapeutics. Compared to the putative genes encrypted in the genome, AMPs that are produced under stress are active forms with the ability to combat resistant microbial species. Within this study, plant-derived AMPs, namely, defensins, nodule-specific cysteine-rich peptides, snakins, lipid transfer proteins, hevein-like proteins, α-hairpinins, and aracins, expressed under biotic and abiotic stresses, are classified. We could observe that while α-hairpinins and snakins display a helix-turn-helix structure, conserved motif patterns such as β1αβ2β3 and β1β2β3 exist in plant defensins and hevein-like proteins, respectively. According to the co-expression data, several plant AMPs are expressed together to trigger synergistic effects with membrane disruption mechanisms such as toroidal pore, barrel-stave, and carpet models. The application of AMPs as an eco-friendly strategy in maintaining agricultural productivity through the development of transgenes and bio-pesticides is discussed. These AMPs can be consumed in packaging material, wound-dressing products, coating catheters, implants, and allergology. AMPs with cell-penetrating properties are verified for the clearance of intracellular pathogens. Finally, the dominant pharmacological activities of bioactive peptides derived from the gastrointestinal digestion of plant AMPs, namely, inhibitors of renin and angiotensin-converting enzymes, dipeptidyl peptidase IV and α-glucosidase inhibitors, antioxidants, anti-inflammatory, immunomodulating, and hypolipidemic peptides, are analyzed. Conclusively, as phytopathogens and human pathogens can be affected by plant-derived AMPs, they provide a bright perspective in agriculture, breeding, food, cosmetics, and pharmaceutical industries, translated as farm to bedside.

Bacteriotherapy represents an attractive approach for both prophylaxis and treatment of human diseases. However, combining probiotic bacteria in "cocktails" is underexplored, despite its potential as an alternative multi-target therapy. Herein, three-strain probiotic mixtures containing different combinations of Bacillus (Bc.) coagulans [ATB-BCS-042], Levilactobacillus (Lv.) brevis [THT 0303101], Lacticaseibacillus (Lc.) paracasei [THT 031901], Bacillus subtilis subsp. natto [ATB-BSN-049], Enterococcus faecium [ATB-EFM-030], and Bifidobacterium (Bf.) animalis subsp. lactis [THT 010802] were prepared. Four cocktails (PA: Bc. coagulans + Lv. brevis + Lc. paracasei, PB: Bc. subtilis subsp. natto + Lv. brevis + Lc. paracasei, PC: E. faecium + Lv. brevis + Lc. paracasei, PD: Bc. coagulans + Lv. brevis + Bf. animalis subsp. lactis) were tested using a short-term (72 h) simulation of the human colonic microbiota in a final dose of 6 × 10⁹ CFU. All these probiotic mixtures significantly increased butyrate production compared to the parallel control experiment. PA and PB promoted a bifidogenic effect and facilitated lactobacilli colonization. Furthermore, reporter gene assays using the AhR_HT29-Lucia cell line revealed that fermentation supernatants from PA and PB notably induced AhR transactivity. Subsequent examination of the metabolic outputs of PA and PB in intestinal epithelial models using cell culture inserts suggested no significant impact on the transepithelial electrical resistance (TEER). Assessment of the expression of proinflammatory and anti-inflammatory cytokines, as well as AhR-related target genes in the Caco-2 cell monolayers indicated that PB's metabolic output upregulated most of the measured endpoints. This in vitro investigation evaluated the potential impact of four multispecies probiotic mixtures in the human colonic microbiota and identified a promising formulation comprising a combination of Bc. subtilis subsp. natto, Lv. brevis, and Lc. paracasei as a promising formulation for further study.

Whole-cell immobilisation technology involving ℽ-aminobutyric acid GABA biosynthesis using lactic acid bacteria (LAB) has been extensively studied owing to its numerous benefits over free-living bacteria, including enhanced productivity, improved cell viability, ability to prevent cell lysis and protect cells against bacteriophages and other stressful conditions. Therefore, a novel LAB biocatalyst was developed using various fruit and fruit waste, immobilising a potential probiotic strain, Lactiplantibacillus plantarum B7, via an adsorption method to improve GABA and cell viability. Apple and watermelon rind have been known to be the ideal natural supports for L. plantarum B7 owing to higher GABA and lactic acid production and improved cell viability among the other natural supports tested and selected to be used in repeated batch fermentation (RBF) to improve GABA production and cell viability. In general, immobilisation of L. plantarum B7 on natural support has better GABA and lactic acid production with improved cell viability via RBF compared to free cells. Watermelon rind-supported cells and apple-supported cells could produce nine and eight successful GABA cycles, respectively, within RBF, whereas free cells could only produce up to four cycles. When using watermelon rind-supported cells and apple-supported cells in RBF, the GABA titer may be raised by up to 6.7 (218.480 ± 0.280 g/L) and 6 (195.439 ± 0.042 g/L) times, respectively, in comparison to GABA synthesis by free cells in single batch fermentation (32.65 ± 0.029 g/L). Additionally, natural support immobilised L. plantarum B7 could retain half of its cell viability even after the 12th cycle of RBF, while no cell was observed in control.

Numerous recent studies have demonstrated that the commensal microbiota plays an important role in host immunity against infections. During the infection process, viruses can exhibit substantial and close interactions with the commensal microbiota. However, the associated mechanism remains largely unknown. Therefore, in this study, we explored the specific mechanisms by which the commensal microbiota modulates host immunity against viral infections. We found that the expression levels of type I interferon (IFN-I) and antiviral priming were significantly downregulated following the depletion of the commensal microbiota due to treatment with broad-spectrum antibiotics (ABX). In addition, we confirmed a unique molecular mechanism underlying the induction of IFN-I mediated by the commensal microbiota. In vivo and in vitro experiments confirmed that Lactobacillus rhamnosus GG (LGG) can suppress herpes simplex virus type 2 (HSV-2) infection by inducing IFN-I expression via the retinoic acid-inducible gene-I (RIG-I) signalling pathway. Therefore, the commensal microbiota-induced production of IFN-I provides a potential therapeutic approach to combat viral infections. Altogether, understanding the complexity and the molecular aspects linking the commensal microbiota to health will help provide the basis for novel therapies already being developed.

Probiotics are widely recognized for their ability to prevent and therapy antibiotic-associated diarrhea (AAD). This study was designed to evaluate Lactiplantibacillus plantarum ELF051 ability to prevent colon inflammation and its effect on gut microbial composition in a mouse model of AAD. The mice were intragastrically administered triple antibiotics for 7 days and then subjected to L. plantarum ELF051 for 14 days. The administration of L. plantarum ELF051 ameliorated the pathological changes in the colon tissue, downregulated interleukin (IL)-1β and tumor necrosis factor (TNF)-α, and upregulated IL-10, and increased the intestinal short-chain fatty acids (SCFAs) level. Lactiplantibacillus plantarum ELF051 also regulated the Toll-like receptor/myeloid differentiation primary response 88/nuclear factor kappa light chain enhancer of activated B cells (TLR4/MyD88/NF-κB) and the phosphatidylinositol 3-kinase/protein kinase B/ NF-κB (PI3K/AKT/ NF-κB) inflammatory signaling pathways. 16S rRNA analyses showed that L. plantarum ELF051 increased the abundance and diversity of gut bacteria, restoring gut microbiota imbalance. A Spearman's rank correlation analysis showed that lactobacilli are closely associated with inflammatory markers and SCFAs. This work demonstrated that L. plantarum ELF051 can attenuate antibiotic-induced intestinal inflammation in a mouse AAD model by suppressing the pro-inflammatory response and modulating the gut microbiota.